title: Staphylococcal SSL5 inhibits leukocyte activation by chemokines and anaphylatoxins.
abstract: Staphylococcus aureus secretes several virulence factors modulating immune responses. Staphylococcal superantigen-like (SSL) proteins are a family of 14 exotoxins with homology to superantigens, but with generally unknown function. Recently, we showed that SSL5 binds to P-selectin glycoprotein ligand 1 dependently of sialyl Lewis X and inhibits P-selectin-dependent neutrophil rolling. Here, we show that SSL5 potently and specifically inhibits leukocyte activation by anaphylatoxins and all classes of chemokines. SSL5 inhibited calcium mobilization, actin polymerization, and chemotaxis induced by chemokines and anaphylatoxins but not by other chemoattractants. Antibody competition experiments showed that SSL5 targets several chemokine and anaphylatoxin receptors. In addition, transfection studies showed that SSL5 binds glycosylated N-termini of all G protein-coupled receptors (GPCRs) but only inhibits stimuli of protein nature that require the receptor N-terminus for activation. Furthermore, SSL5 increased binding of chemokines to cells independent of chemokine receptors through their common glycosaminoglycan-binding site. Importance of glycans was shown for both GPCR and chemokine binding. Thus, SSL5 is an important immunomodulatory protein of S aureus that targets several crucial, initial stages of leukocyte extravasation. It is therefore a potential new antiinflammatory compound for diseases associated with chemoattractants and their receptors and disorders characterized by excessive recruitment of leukocytes.
authors.affiliation: Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands. j.bestebroer@umcutrecht.nl
authors.forename: Jovanka
authors.initials: J
authors.lastname: Bestebroer
authors.forename: Kok P M
authors.initials: KP
authors.lastname: van Kessel
authors.forename: Hafida
authors.initials: H
authors.lastname: Azouagh
authors.forename: Annemiek M
authors.initials: AM
authors.lastname: Walenkamp
authors.forename: Ingrid G J
authors.initials: IG
authors.lastname: Boer
authors.forename: Roland A
authors.initials: RA
authors.lastname: Romijn
authors.forename: Jos A G
authors.initials: JA
authors.lastname: van Strijp
authors.forename: Carla J C
authors.initials: CJ
authors.lastname: de Haas
mesh_terms: D000706:Anaphylatoxins / Q000276:immunology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000276:immunology* / Q000378:metabolism; D020013:Calcium Signaling / Q000276:immunology; D002460:Cell Line; D018925:Chemokines / Q000276:immunology / Q000378:metabolism; D005098:Exotoxins / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007155:Immunologic Factors / Q000276:immunology* / Q000378:metabolism; D036904:Leukocyte Rolling / Q000276:immunology*; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D018375:Neutrophil Activation / Q000276:immunology*; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding / Q000276:immunology; D019707:Receptors, Chemokine / Q000276:immunology / Q000378:metabolism; D013211:Staphylococcus aureus / Q000276:immunology* / Q000378:metabolism / Q000472:pathogenicity; D014312:Trisaccharides / Q000276:immunology / Q000378:metabolism; D037521:Virulence Factors / Q000276:immunology* / Q000378:metabolism
chemical_list: D000706:Anaphylatoxins; D001426:Bacterial Proteins; D018925:Chemokines; D005098:Exotoxins; D007155:Immunologic Factors; D016256:Lewis X Antigen; D008562:Membrane Glycoproteins; C084882:P-selectin ligand protein; D019707:Receptors, Chemokine; D014312:Trisaccharides; D037521:Virulence Factors; C110530:galactosyl-(1,4)-fucopyranosyl-(1,3)-N-acetylglucosamine